Dear all,
I had a problem with transfected S2 cell. I observed that many cells were
going to die when I tried to expand them after 3 weeks of selection of
hygromicin-B resistance.
Situations are as follows:
I have introduced pCoHygro and pMT-HisV5A with a gene of my interest by
CaCl2 method into drosophila S2 cells. These cells are cultured for 3 weeks
in the medium of M3/10%FCS with 300ug/ml hygromicin-B that were replaced
every 4 or 5 days. During that period, there is no cell debris(, although
cells with a bigger mass were observed). After that, cells were reached a
density of 3.3x10^6 cells/ml and passed at 1:2 dilution into an old and new
flask. Two days later, a lot of cell debris were obserbed in both flasks.
Should I pass them with a smaller volume?
Should I use old medium to support their growth?
Any comments are welcome.
Sincerely,
Jun-ichi
-- Jun-ichi Aikawa, Ph.D.
Cellular Biochemistry Laboratory, RIKEN
2-1 Hirosawa, Wako-shi, Saitama 351-0198, JAPAN
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